Xenbase Image ID: 41385

Figure 5. Inhibition of ETV2 function strongly reduces endothelial, but not hematopoietic marker expression. A,B: Antisense etv2 morpholino oligomer 1 (MO1) efficiently inhibits enhanced green fluorescent protein (EGFP) expression from a target transcript in control experiments. A: Brightfield and fluorescent image of embryos injected with etv2-EGFP fusion transcript and etv2 mismatch control MO (CoMO), showing strong EGFP fluorescence. B: Brightfield and fluorescent image of embryos injected with etv2-EGFP fusion transcript and etv2 MO1. EGFP fluorescence is almost eliminated. C-M: Embryos injected with control MO (CoMo) or etv2 MO1 and assayed for marker gene expression (as indicated) by in situ hybridization. All embryos were assayed at approximately stage 32. Inset boxes in panels C-H, show slightly enlarged and/or increased contrast views centered on the intersegmental vessels and posterior cardinal vein. C-H: Treatment with etv2 MO1 effectively eliminated expression of the endothelial marker genes flk1, aplnr, and erg. In H, expression of erg in neural crest cells in the branchial arch region (open arrowhead) is unchanged. I: etv2 MO1 knockdown of endothelial marker expression is efficiently rescued with etv2 mRNA. All embryos were injected with 50 ng of MO1 plus varying amounts of etv2 mRNA as indicated. Marker gene expression was assayed by in situ hybridization and embryos showing mostly normal endothelial pattern were scored as wild-type. The number in each bar indicates the number of embryos scored for each treatment. Asterisks show statistically significant differences relative to value of MO1 alone, analyzed using Chi-squared test. J-O: Treatment with etv2 MO1 had no detectable effect on expression of myeloid (runx1 and spib) or erythroid (tadpole alpha -globin) markers. Image published in: Salanga MC et al. (2010) Copyright © 2010. Image reproduced with permission of the Publisher, John Wiley & Sons. Gene Synonyms Species Stage(s) Tissue spib.L spi-b, spib-a, spib-b X. laevis Throughout NF stage 29 and 30 to NF stage 32 ventral blood island blood vessel runx1.L aml, aml-1, aml1, aml1 oncogene, aml1-evi-1, amlcr1, cbfa2, evi-1, LOC108709238, pebp2ab, Runx-1, Xaml, Xaml1 X. laevis Throughout NF stage 32 ventral blood island olfactory region periotic region kdr.L flk-1, flk1, LOC108698463, vegfr-2, vegfr2, xflk-1 X. laevis Throughout NF stage 32 branchial arch posterior cardinal vein vascular plexus erg.L erg-3, erg-a, erg-b, MGC80765, p55, vascular ETS factor X. laevis Throughout NF stage 32 to NF stage 33 and 34 posterior cardinal vein vascular plexus branchial arch ophthalmic artery anterior cardinal vein intersomitic vessel hba1.S alpha globin, alpha-globin, T3 globin X. laevis Throughout NF stage 32 ventral blood island aplnr.L agtrl1, angio1, apj, apjr, aplnr-a, aplnr-b, hg11, msr, X-msr, Xangio1, Xmsr X. laevis Throughout NF stage 33 and 34 branchial arch vascular plexus intersomitic vessel posterior cardinal vein ophthalmic artery tail bud duct of Cuvier Image source: Published Larger Image Printer Friendly View

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